US 11,713,471 B2
Class II, type V CRISPR systems
Brian Thomas, Emeryville, CA (US); Christopher Brown, Emeryville, CA (US); Audra Devoto, Emeryville, CA (US); Cristina Butterfield, Emeryville, CA (US); Lisa Alexander, Emeryville, CA (US); Daniela S. A. Goltsman, Emeryville, CA (US); Justine Albers, Emeryville, CA (US); Alan Brooks, Emeryville, CA (US); Greg Cost, Emeryville, CA (US); Morayma Temoche-Diaz, Emeryville, CA (US); Cindy Castelle, Emeryville, CA (US); and Rebecca Lamothe, Emeryville, CA (US)
Assigned to METAGENOMI, INC., Emeryville, CA (US)
Filed by METAGENOMI, INC., Emeryville, CA (US)
Filed on Mar. 28, 2022, as Appl. No. 17/706,466.
Application 17/706,466 is a continuation of application No. PCT/US2021/021259, filed on Mar. 6, 2021.
Claims priority of provisional application 63/116,157, filed on Nov. 19, 2020.
Claims priority of provisional application 63/069,699, filed on Aug. 24, 2020.
Claims priority of provisional application 63/068,316, filed on Aug. 20, 2020.
Claims priority of provisional application 63/045,815, filed on Jun. 29, 2020.
Claims priority of provisional application 63/022,276, filed on May 8, 2020.
Claims priority of provisional application 62/986,477, filed on Mar. 6, 2020.
Prior Publication US 2022/0290187 A1, Sep. 15, 2022
Int. Cl. C12N 15/90 (2006.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01)
CPC C12N 15/90 (2013.01) [C12N 9/22 (2013.01); C12N 15/1068 (2013.01); C12N 15/11 (2013.01); C12N 15/111 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 26 Claims
 
1. An engineered nuclease system comprising:
a. an endonuclease configured to be selective for a protospacer adjacent motif (PAM) sequence comprising the sequence 5′-YYn-3′ (SEQ ID NO: 3871), wherein said endonuclease is a class 2, type V Cas endonuclease, wherein said endonuclease comprises a WED II domain and a PAM-interacting domain having at least 80% amino acid sequence identity to the WED II domain and the PAM-interacting domain of the amino acid sequence of SEQ ID NO: 215; and
b. an engineered guide RNA, wherein said engineered guide RNA is configured to form a complex with said endonuclease and said engineered guide RNA comprises a spacer sequence configured to hybridize to a target nucleotide sequence.